沉默表皮生长因子受体的表达对肺腺癌A549细胞自噬的影响

doi:10.3969/j.issn.1004-616x.2019.05.002

Abstract

Abstract: OBJECTIVE:The role of EGFR in autophagy activity in lung adenocarcinoma A549 cells was investigated using small interference RNA to silence EGFR expression. METHODS:A549 cells were divided into four groups:untreated,EGFR siRNA-1,EGFR siRNA-2 and EGFR siRNA-3 groups. Real-time RT-PCR and Western blot were used to detect degradation of EGFR after transfection with EGFR siRNA. In addition,A549 cells were treated with different EGF concentrations:0,10,25 and 50 ng/mL. Protein expression of EGFR was detected using Western blot. Furthermore,A549 cells were divided into four groups:EGFR siRNA,negative control siRNA,transfection reagent (only add equivalent transfection reagent) and untreated groups. All four groups also received 50 ng/mL EGF. Western blot was used to detect expression of LC3Ⅱ/LC3Ⅰ which was observed as the effect on silencing EGFR expression on autophagy activity. A549 cells were divided into EGFR siRNA transfection and negative control siRNA groups,and autophagosomes were observed by transmission electron microscope. RESULTS:After treatment of A549 cells with the three different EGFR siRNAs,the mRNA and protein expression of EGFR were significant reduced. Western blot indicates that increased LC3-Ⅱ/LC3-Ⅰ ratios were induced by down-regulation of EGFR. Transmission electron microscopy shows that EGFR siRNA treatment,compared with control siRNA treatment,induced marked formation of autophagsomes. CONCLUSION:Reducing the expression of EGFR increased autophagy activities of lung adenocarcinoma A549 cells.

Key words: EGFR, lung adenocarcinoma, autophagy, siRNA, LC3-Ⅱ/LC3-Ⅰ

CLC Number:

R730.2

MA Yang, WU Juan, JI Xiaokun, WANG Rui, DU Yun, WANG Heng, GUO Xiao, ZHANG Yan. Silencing EGFR expression influenced autophagy activity in A549 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2019, 31(5): 347-351,358.

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Silencing EGFR expression influenced autophagy activity in A549 cells

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